Construction and Expression of Recombinant Adenovirus Encoding Tumor Suppressor Fhit and Its Biological Function in Colon Cancer Cells / 中国生物工程杂志
China Biotechnology
;
(12)2006.
Article
in Chinese
| WPRIM
| ID: wpr-596865
ABSTRACT
AIM:
To construct a recombinant adenovirus carrying Fhit gene,a tumor suppressor in many types of cancer,and to observe its biological function on the proliferation of colon cancer cells.METHODS:
Fhit gene was cloned from the fetal liver cDNA library using the PCR method.The PCR product was inserted into the T vector to construct the plasmid pMD18T-Fhit.The Fhit fragment from the pMD18T-Fhit was inserted into the vector ptrack-CMV to construct a shuttle plasmid ptrack-CMV-Fhit.After PmeI digested and linearized process,ptrack-CMV-Fhit was co-transformed into Escherichia coli strain BJ5183 together with the adenovirus backbone vector pAdEasy-1 to generate a recombinant adenovirus plasmid by homologous recombination.The adenovirus plasmid was identified by PacI digestion and transfected into 293A cells to package a recombinant adenovirus which expressed the Fhit protein.Furthermore,the adenovirus rAd-Fhit was infected into colon cancer cells,and the expression of the ectogenic protein was detected by Western blotting.Finally,the proliferation of colon cancer cells was observed in adenovirus-infected cells by the MTT assay.RESULTS:
Constructed the recombinant adenovirus encoding Fhit gene and expressed it in colon cancer cells successfully.Detected that the proliferation of colon cancer cells was inhibited obviously in rAd-Fhit-infected cells with comparison to the control groups.CONCLUSION:
Fhit may function as a tumor suppressor in colon cancer cells,and the adenovirus-mediated Fhit can be a novel strategy for the colon cancer therapeutics.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
China Biotechnology
Year:
2006
Type:
Article
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